Hydrogel for cell-laden bioprinting, bioink, and preparation method and application thereof

Abstract

A hydrogel for cell-laden bioprinting, bioink, and a preparation method and an application thereof, relates to the technical field of biomedical polymer hydrogels. The hydrogel for cell-laden bioprinting is polymer gel formed by adding a cell-specific material into a matrix of alginate and gelatin and crosslinking and curing, wherein the cell-specific material is polypeptide selected according to different laden cells. The structures printed using the hydrogel may have the advantages such as adjustable mechanical properties, adjustable porosity, high biocompatibility, high printing accuracy, and high customizability, which may widely support the printing of human tissues and organs such as spinal cord, cartilage, and heart, and has good prospects for applications in tissue repair, organ transplantation and so on.

What is claimed is: 
     
       1. A hydrogel for cell-laden bioprinting comprising a polymer gel formed by adding a polypeptide cell-specific material into a matrix of alginate and gelatin and crosslinking and curing the matrix. 
     
     
       2. A method of utilizing the hydrogel for cell-laden bioprinting of  claim 1  comprising applying the hydrogel for cell-laden bioprinting in the field of biomedicine. 
     
     
       3. A method for preparing a cell-laden bioink, comprising the steps of:
 preparing a cell-laden bioink precursor via adding sodium alginate to phosphate buffered saline (PBS) and dissolving by stirring and warming thereby obtaining an alginate solution, 
 adding and dissolving gelatin and a polypeptide cell-specific material to the PBS thereby obtaining a gelatin composite solution, 
 mixing the alginate solution and the gelatin composite solution evenly thereby obtaining a bioink solution, and 
 centrifuging and adding a suspension of cells into the bioink solution and blowing the cells to make the cells disperse and even thereby obtaining the cell-laden bioink. 
 
     
     
       4. The method for preparing the cell-laden bioink according to  claim 3 , wherein a concentration range of the alginate solution is 2% to 5% (w/v) and a concentration range of the gelatin solution is 2% to 8% (w/v). 
     
     
       5. The method for preparing the cell-laden bioink according to  claim 3 , wherein the cells are L929 cells, the polypeptide cell-specific material is CMP27, the concentration of the alginate solution is 2.5% (w/v) and the concentration of the gelatin solution is 6% (w/v). 
     
     
       6. A method for preparing a cell-laden scaffold, comprising the steps of:
 loading a cell-laden bioink prepared by the preparation method of  claim 3  into a syringe of a 3D bio-printer, 
 extruding the cell-laden bioink onto a cryogenic platform through a pneumatic nozzle on the 3D bio-printer thereby preparing a grid-like cell-laden scaffold, 
 soaking the printed cell-laden scaffold in a 4.5% (w/v) calcium chloride solution for 5 minutes, and 
 washing with PBS thereby obtaining a final structure of the cell-laden scaffold. 
 
     
     
       7. A cell-laden scaffold prepared by the method for preparing the cell-laden scaffold of  claim 6 . 
     
     
       8. A method of utilizing the cell-laden scaffold of  claim 7  comprising applying the cell- laden scaffold in the field of biomedicine. 
     
     
       9. An adhesive bandage, comprising the cell-laden scaffold of  claim 7 . 
     
     
       10. A method for preparing tissues and organs utilizing a freeform reversible embedding of suspended hydrogels (FRESH) extrusion printing, comprising the steps of:
 fixing a gelatin support bath on a cryogenic platform, 
 loading a cell-laden bioink prepared by the preparation method of  claim 3  into a syringe of a 3D bio-printer, 
 extruding and printing the cell-laden bioink into the gelatin support bath through a pneumatic nozzle, 
 raising a temperature of the cryogenic platform after, 
 releasing the printed structure, and 
 washing the printed structure with PBS. 
 
     
     
       11. The method for preparing tissues and organs utilizing the FRESH extrusion printing according to  claim 10 , further comprising the steps of:
 incubating a prepared solution containing 5% (w/v) gelatin and 1% (w/v) CaCl 2  at 4° C. for 24 hours, 
 crushing the incubated solution utilizing a crusher after adding 3 times the volume of the CaCl 2  solution thereby obtaining gelatin granules; 
 centrifuging the gelatin granules, 
 removing the supernatant, 
 washing 3 times with the 1% (w/v) CaCl 2  solution, 
 centrifuging the gelatin granules again thereby obtaining a gelatin slurry, and transferring the gelatin slurry to a petri dish. 
 
     
     
       12. A method for preparing a cell-laden scaffold, comprising the steps of:
 loading the cell-laden bioink prepared by the preparation method of  claim 4  into a into a syringe of a 3D bio-printer, 
 extruding the cell-laden bioink onto a cryogenic platform through a pneumatic nozzle on the 3D bio-printer thereby preparing a grid-like cell-laden scaffold, 
 soaking the printed cell-laden scaffold in a 4.5% (w/v) calcium chloride solution for 5 minutes, and 
 washing with PBS thereby obtaining a final structure of the cell-laden scaffold. 
 
     
     
       13. A cell-laden scaffold comprising the cell-laden scaffold prepared by the preparation method of  claim 12 . 
     
     
       14. An adhesive bandage, comprising the cell-laden scaffold of  claim 13 . 
     
     
       15. A method of utilizing the cell-laden scaffold of  claim 13  comprising applying the cell-laden scaffold in the field of biomedicine. 
     
     
       16. A method for preparing a cell-laden scaffold, comprising the steps of:
 loading the cell-laden bioink prepared by the preparation method of  claim 5  into a into a syringe of a 3D bio-printer, 
 extruding the cell-laden bioink onto a cryogenic platform through a pneumatic nozzle on the 3D bio-printer thereby preparing a grid-like cell-laden scaffold, 
 soaking the printed cell-laden scaffold in a 4.5% (w/v) calcium chloride solution for 5 minutes, and 
 washing with PBS thereby obtaining a final structure of the cell-laden scaffold. 
 
     
     
       17. A cell-laden scaffold comprising the cell-laden scaffold prepared by the preparation method of  claim 16 . 
     
     
       18. An adhesive bandage, comprising the cell-laden scaffold of  claim 17 . 
     
     
       19. A method of utilizing the cell-laden scaffold of  claim 17  comprising applying the cell-laden scaffold in the field of biomedicine.